The Characterization of GSDMB Splicing and Backsplicing Profiles Identifies Novel Isoforms and a Circular RNA That Are Dysregulated in Multiple Sclerosis.
The Characterization of GSDMB Splicing and Backsplicing Profiles Identifies Novel Isoforms and a Circular RNA That Are Dysregulated in Multiple Sclerosis.
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DOI：
10.3390/ijms18030576
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发表时间：
2017-03-07
影响因子：
5.6
通讯作者：
Asselta R
中科院分区：
文献类型：
--
作者：
Cardamone G;Paraboschi EM;Rimoldi V;Duga S;Soldà G;Asselta R
选择性剪接（alternative splicing，AS）异常是自身免疫性疾病（autoimmune diseases，AIDS）的常见特征.特别是，越来越多的证据表明，剪接调节基因的普遍缺陷和多发性硬化症（MS）之间存在致病关联。此外，几项研究已经证明了MS患者中选择性剪接异构体的不平衡可能有助于疾病病因。在这项工作中，使用基于PCR的技术（逆转录（RT）-PCR，荧光竞争，实时，和数字RT-PCR检测）的组合，我们调查了选择性剪接基因编码Gasdermin B，GSDMB，这是反复与哮喘和艾滋病的易感性。GSDMB AS和反向剪接谱的深入表征使我们鉴定了外显子环状RNA（ecircRNA）以及新型GSDMB框内和框外同种型。在人类细胞系中，非生产性剪接变体被无义介导的mRNA衰减（NMD）下调，表明GSDMB水平被NMD显著调节。重要的是，与对照组相比，AS亚型和鉴定的ecircRNA在复发缓解型MS患者的外周血单核细胞中显著失调，进一步支持异常RNA代谢是该疾病的特征性特征的观点。
Abnormalities in alternative splicing (AS) are emerging as recurrent features in autoimmune diseases (AIDs). In particular, a growing body of evidence suggests the existence of a pathogenic association between a generalized defect in splicing regulatory genes and multiple sclerosis (MS). Moreover, several studies have documented an unbalance in alternatively-spliced isoforms in MS patients possibly contributing to the disease etiology. In this work, using a combination of PCR-based techniques (reverse-transcription (RT)-PCR, fluorescent-competitive, real-time, and digital RT-PCR assays), we investigated the alternatively-spliced gene encoding Gasdermin B, GSDMB, which was repeatedly associated with susceptibility to asthma and AIDs. The in-depth characterization of GSDMB AS and backsplicing profiles led us to the identification of an exonic circular RNA (ecircRNA) as well as of novel GSDMB in-frame and out-of-frame isoforms. The non-productive splicing variants were shown to be downregulated by the nonsense-mediated mRNA decay (NMD) in human cell lines, suggesting that GSDMB levels are significantly modulated by NMD. Importantly, both AS isoforms and the identified ecircRNA were significantly dysregulated in peripheral blood mononuclear cells of relapsing-remitting MS patients compared to controls, further supporting the notion that aberrant RNA metabolism is a characteristic feature of the disease.